LOCUS       sequence-488204-        7332 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_185, derived from human fusion protein SSR1_FARS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7332)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7332)
  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_185
FEATURES             Location/Qualifiers
     source          1..7332
                     /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(865..1575,1576..1578,1579..1581))
                     /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(907..928)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1256..1275
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1515..1536
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1578..1587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1663..1866)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1692..1712)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1867..2170
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2427..2471)
                     /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    2656..2889
                     /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    3249..3366
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3858..3983
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4030..4287
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4289..4668
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4699..4803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4804..4872,4873..5664)
                     /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(5022..5041)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5835..6423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6324..6343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6577..6594
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6662..6796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6686..6705)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6740..6759
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6802..6997)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6816..6951
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7056..7077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7092..7097,7098..7115,7116..7122)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7130..7146
                     /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     7135..7157
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7154..7170
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7154..7170
                     /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 gtctgcagat ggactagctt ctggttctcc
      601 ggaaacgtct tcttcttctt tatcttctac gaggtcagta ggttcgtctt cctcaacctc
      661 ggcttcatcg tcctcgtctt caataatgct gtcttcgacc gtctcctcgt cctcagtgag
      721 atcctgagct accgccagaa ggcctctagg tccgccccta aacagaaccg tagccggaaa
      781 caccaacaac agaaggagga ggagccgggg caacagtctc atccgggaag cggccgcact
      841 tcccccggag cctccgcttc cgcccttgta cagctcgtcc atgccgagag tgatcccggc
      901 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
      961 cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     1021 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     1081 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     1141 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     1201 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     1261 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     1321 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     1381 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     1441 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     1501 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     1561 ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga
     1621 ctgcagaatt cggttcacta aacgagcccg gttcactaaa cgagctctgc ttatatagac
     1681 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     1741 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     1801 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     1861 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     1921 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     1981 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     2041 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     2101 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     2161 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     2221 ttaataacta gcacgcgttc gaagcttgag ctcggtcccc tcggggttgg gaggtgggtc
     2281 tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag tacagcaaaa
     2341 actattctta aacctaccaa gcctcctact atcattatga ataattttat ataccacagc
     2401 caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc caataattct
     2461 tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat taagcttgtg
     2521 taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat ctgttccaga
     2581 gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag ttttccagag
     2641 caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc caggattctt
     2701 gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg cgcctcaata
     2761 gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct ggcctgtacc
     2821 gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa cccaaggaac
     2881 aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt tgccttggtg
     2941 ggtgctactc ctaatggttc aatttttact actttatatt tatataattc acttctccaa
     3001 ttgtccctca tatctcctcc ttatggtgtt ttactaaact tttccatgtt ctaatcctca
     3061 tcctgtctac ttgccacaca atcatcacct gccatctgtt ttccataatc cctaatgatc
     3121 tttgcttttc ttcttggcac tacttttatg tcactattat cttgtattac tactgcccct
     3181 tcacctttcc agaggagctt tgctggtcct ttccaaagtg gatttctgct gtccctgtaa
     3241 taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct
     3301 gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt
     3361 ttaaaattgt ggatgaatac tgccatttgt actgctgtct taagatgttc agcctgatct
     3421 cttacctgtc ctataatttt ctttaattct ttattcatag attctactac tccttgactt
     3481 tggggattgt agggaatttc caggtctgaa gatcctgctg cttgctgtgc ttttttctta
     3541 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     3601 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     3661 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     3721 aacaggccag tattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     3781 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     3841 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     3901 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     3961 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     4021 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     4081 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     4141 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     4201 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     4261 tccccagtcc cgcccaggcc acgcctccca tggtaatagc gatgactaat acgtagatgt
     4321 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     4381 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     4441 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     4501 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     4561 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     4621 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga cccaggtggc
     4681 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4741 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4801 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4861 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4921 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4981 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5041 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5101 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5161 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5221 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5281 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5341 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5401 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5461 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5521 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5581 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5641 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5701 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5761 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5821 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5881 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5941 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6001 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6061 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6121 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6181 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6241 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6301 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6361 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6421 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6481 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6541 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6601 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6661 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     6721 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     6781 ctgcaataaa caagttagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     6841 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     6901 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     6961 taggggcggg actatggttg ctgactaatt gagatgcgga tccgctggca cgacaggttt
     7021 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7081 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7141 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgg gctgcaggtc
     7201 gagggatctc cataagagaa gagggacagc tatgactggg agtagtcagg agaggaggaa
     7261 aaatctggct agtaaaacat gtaaggaaaa ttttagggat gttaaagaaa aaaataacac
     7321 aaaacaaaat at
//
