LOCUS       sequence-488200-        7281 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_180, derived from human fusion protein SNTB2_ZCRB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7281)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7281)
  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_180
FEATURES             Location/Qualifiers
     source          1..7281
                     /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(814..1524,1525..1527,1528..1530))
                     /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(856..877)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1205..1224
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1464..1485
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1527..1536
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1612..1815)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1641..1661)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1816..2119
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2376..2420)
                     /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    2605..2838
                     /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    3198..3315
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3807..3932
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3979..4236
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4238..4617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4648..4752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4753..4821,4822..5613)
                     /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(4971..4990)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5784..6372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6273..6292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6526..6543
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6611..6745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6635..6654)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6689..6708
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6751..6946)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6765..6900
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7005..7026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7041..7046,7047..7064,7065..7071)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7079..7095
                     /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     7084..7106
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7103..7119
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7103..7119
                     /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 tacacggaca cggcgaactg gggggctcgc
      601 ccccgcctca ccggcggggc cacggggcgg agcgggcggt gacggtggac ccaatccgcg
      661 agagggcgaa ccgggtaagc tatccccagc gccgccacca ttgggcagcc cattgaaagc
      721 tgcagccgcg ggaccaagag cgggctccag ttctgccgcg gctgcgtcac cccgggaagc
      781 ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca tgccgagagt
      841 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
      901 tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
      961 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     1021 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     1081 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     1141 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     1201 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     1261 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     1321 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     1381 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     1441 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     1501 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg
     1561 taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac gagctctgct
     1621 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     1681 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     1741 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     1801 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     1861 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     1921 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     1981 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     2041 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     2101 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     2161 tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct cggggttggg
     2221 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     2281 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     2341 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     2401 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     2461 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     2521 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     2581 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     2641 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     2701 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     2761 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     2821 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     2881 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     2941 cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt ttccatgttc
     3001 taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt tccataatcc
     3061 ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc ttgtattact
     3121 actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg atttctgctg
     3181 tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt
     3241 tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc
     3301 ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt aagatgttca
     3361 gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga ttctactact
     3421 ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc ttgctgtgct
     3481 tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     3541 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     3601 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     3661 gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct tgcccatact
     3721 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     3781 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     3841 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     3901 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     3961 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     4021 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     4081 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     4141 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
     4201 gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg atgactaata
     4261 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4321 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4381 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4441 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4501 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4561 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcgac
     4621 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4681 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4741 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4801 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4861 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4921 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4981 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5041 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5101 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5161 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5221 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5281 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5341 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5401 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5461 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5521 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5581 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5641 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5701 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5761 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5821 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5881 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5941 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6001 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6061 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6121 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6181 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6241 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6301 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6361 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6421 tttgctcaca tgttctttcc tgcgttatcc cctgcttctg tggataaccg tattaccgcc
     6481 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6541 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6601 taatgcagcg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     6661 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6721 cattataagc tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc
     6781 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     6841 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     6901 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
     6961 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7021 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7081 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     7141 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     7201 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     7261 aaataacaca aaacaaaata t
//
