LOCUS       sequence-488229-        7422 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_214, derived from human fusion protein ZFAND3_DGKH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7422)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7422)
  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_214
FEATURES             Location/Qualifiers
     source          1..7422
                     /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(955..1665,1666..1668,1669..1671))
                     /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(997..1018)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1346..1365
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1605..1626
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1668..1677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1753..1956)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1782..1802)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1957..2260
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2517..2561)
                     /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    2746..2979
                     /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    3339..3456
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3948..4073
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4120..4377
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4379..4758
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4789..4893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4894..4962,4963..5754)
                     /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(5112..5131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5925..6513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6414..6433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6667..6684
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6752..6886
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6776..6795)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6830..6849
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6892..7087)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6906..7041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7146..7167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7182..7187,7188..7205,7206..7212)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7220..7236
                     /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     7225..7247
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7244..7260
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7244..7260
                     /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 ggacttttgc ttcgagcgac tggtctcctc
      601 gctacgttcg gtgttctcca gtaaacgggg gcgttttacg gggctcgctt cattttccga
      661 ttggctgtcc gtgccgcacg agcgtttggt gggcgtgata agggatacgt gggctgtgtc
      721 cgtgcacggc ccgcattctt ctttgcttgg actcgtgacg ttaagctctg taggtaaggg
      781 ctgttgactt ggagacaaag taggtgtggt gatggacgtg ttattgttat cactagtagt
      841 ttcctcgctg aaaaggtcgg actgcgaatt ggaagttgaa ggtgcgctgt catcgtccgg
      901 ttgcttcttc tgccgggaag cggccgcact tcccccggag cctccgcttc cgcccttgta
      961 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
     1021 atcgcgcttc tcgttggggt ctttgctcag cttggactgg gtgctcaggt agtggttgtc
     1081 gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg
     1141 cacgctgccg tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg
     1201 cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag
     1261 gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc
     1321 gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt
     1381 gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg
     1441 gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca
     1501 gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc
     1561 atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc
     1621 gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcgac
     1681 cggtggatcc cgggcccgcg gtaccgtcga ctgcagaatt cggttcacta aacgagcccg
     1741 gttcactaaa cgagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
     1801 cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
     1861 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     1921 tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac
     1981 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     2041 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     2101 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     2161 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     2221 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     2281 atgaactaat gaccccgtaa ttgattacta ttaataacta gcacgcgttc gaagcttgag
     2341 ctcggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata tccctgccta
     2401 actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa gcctcctact
     2461 atcattatga ataattttat ataccacagc caatttgtta tgttaaacca attccacaaa
     2521 cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg gttttgcgat
     2581 tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc ccactccatc
     2641 caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc attccaaggc
     2701 acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag ctgttgatcc
     2761 tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc ccagactgtg
     2821 agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg ctgctgcact
     2881 ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg
     2941 cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc tcttttttct
     3001 ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc aatttttact
     3061 actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc ttatggtgtt
     3121 ttactaaact tttccatgtt ctaatcctca tcctgtctac ttgccacaca atcatcacct
     3181 gccatctgtt ttccataatc cctaatgatc tttgcttttc ttcttggcac tacttttatg
     3241 tcactattat cttgtattac tactgcccct tcacctttcc agaggagctt tgctggtcct
     3301 ttccaaagtg gatttctgct gtccctgtaa taaacccgaa aattttgaat ttttgtaatt
     3361 tgtttttgta attctttagt ttgtatgtct gttgctatta tgtctactat tctttcccct
     3421 gcactgtacc ccccaatccc cccttttctt ttaaaattgt ggatgaatac tgccatttgt
     3481 actgctgtct taagatgttc agcctgatct cttacctgtc ctataatttt ctttaattct
     3541 ttattcatag attctactac tccttgactt tggggattgt agggaatttc caggtctgaa
     3601 gatcctgctg cttgctgtgc ttttttctta cttttgtttt gctcttcctc tatcttgtct
     3661 aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact
     3721 gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc
     3781 agtatttgtc tacagccttc tgatgtttct aacaggccag tattaactgc gaatcgttct
     3841 agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt
     3901 aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac
     3961 ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc
     4021 ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct
     4081 ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg
     4141 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta
     4201 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag
     4261 agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca aaaagcagct
     4321 gcttatatgc aggatctgag ggctcgccac tccccagtcc cgcccaggcc acgcctccca
     4381 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     4441 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     4501 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     4561 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     4621 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     4681 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     4741 tagtcaataa tcaatgtcga cccaggtggc acttttcggg gaaatgtgcg cggaacccct
     4801 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4861 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4921 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4981 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5041 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5101 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5161 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5221 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5281 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5341 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5401 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5461 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5521 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5581 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5641 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5701 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5761 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5821 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5881 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5941 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6001 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6061 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6121 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6181 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6241 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6301 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6361 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6421 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6481 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6541 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgcttct
     6601 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6661 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6721 cccgcgcgtt ggccgattca ttaatgcagc ggatccagac atgataagat acattgatga
     6781 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     6841 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttagct ttttgcaaaa
     6901 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     6961 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     7021 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     7081 gagatgcgga tccgctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     7141 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7201 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     7261 catgattacg ccaagcttgg gctgcaggtc gagggatctc cataagagaa gagggacagc
     7321 tatgactggg agtagtcagg agaggaggaa aaatctggct agtaaaacat gtaaggaaaa
     7381 ttttagggat gttaaagaaa aaaataacac aaaacaaaat at
//
