LOCUS       sequence-488182-        7752 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_162, derived from human fusion protein RANBP3_SLC25A23.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7752)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7752)
  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_162
FEATURES             Location/Qualifiers
     source          1..7752
                     /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(1285..1995,1996..1998,1999..2001))
                     /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(1327..1348)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1676..1695
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1935..1956
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1998..2007
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2083..2286)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2112..2132)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2287..2590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2847..2891)
                     /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    3076..3309
                     /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    3669..3786
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4278..4403
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4450..4707
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4709..5088
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5119..5223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5224..5292,5293..6084)
                     /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(5442..5461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6255..6843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6744..6763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6997..7014
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7082..7216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7106..7125)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7160..7179
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7222..7417)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7236..7371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7476..7497
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7512..7517,7518..7535,7536..7542)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7550..7566
                     /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     7555..7577
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7574..7590
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7574..7590
                     /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 tgcggcctcc gaggggctgc gccaagccgc
      601 cgtgtcggga cttgcagccg gaacggcccc cgtacaatct gctggcaaag acacgccatt
      661 tgtgccgctg cttggaaccg tttgagacag tgccttaggt tgcggggctt gcagtacggc
      721 tggacgcagg actgagcgtt gctgctcctt cggtttttgc gatggtaacc ctgcagaagg
      781 cgcttggccg tggataaggg ttgggggctt taaacgaaac ccggaactgc gttcctctga
      841 ttgactaggg gggaactgcg tcaatgagga tgtgcgttcg cgtttaacag gtggacaata
      901 atttccgtct tcacggtccg aatcctcccc accctcggga gaactaccgc ctgcgctgcg
      961 cccagctaac tcacggggga aaggaggaag ttgagcctct ggcgcagggg gtggaggtgt
     1021 tgaggctgat gctccagccg gagcaggtgg ttccagggcg tgctcccccg cagattccgg
     1081 gtgtccagta ccgtgatgtg gagcctcagc ctcaccacgt ggctcctctc cactatcgga
     1141 cagatttttt tgttcggcgg gactcttttg acccttatct ttctgaaaca cgaaaaccgg
     1201 gggcgcgatt gcaggttttt cttcgttggc caagtctgcc atccgggaag cggccgcact
     1261 tcccccggag cctccgcttc cgcccttgta cagctcgtcc atgccgagag tgatcccggc
     1321 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     1381 cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     1441 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     1501 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     1561 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     1621 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     1681 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     1741 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     1801 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     1861 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     1921 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     1981 ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga
     2041 ctgcagaatt cggttcacta aacgagcccg gttcactaaa cgagctctgc ttatatagac
     2101 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     2161 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     2221 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     2281 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     2341 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     2401 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     2461 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     2521 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     2581 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     2641 ttaataacta gcacgcgttc gaagcttgag ctcggtcccc tcggggttgg gaggtgggtc
     2701 tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag tacagcaaaa
     2761 actattctta aacctaccaa gcctcctact atcattatga ataattttat ataccacagc
     2821 caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc caataattct
     2881 tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat taagcttgtg
     2941 taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat ctgttccaga
     3001 gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag ttttccagag
     3061 caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc caggattctt
     3121 gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg cgcctcaata
     3181 gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct ggcctgtacc
     3241 gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa cccaaggaac
     3301 aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt tgccttggtg
     3361 ggtgctactc ctaatggttc aatttttact actttatatt tatataattc acttctccaa
     3421 ttgtccctca tatctcctcc ttatggtgtt ttactaaact tttccatgtt ctaatcctca
     3481 tcctgtctac ttgccacaca atcatcacct gccatctgtt ttccataatc cctaatgatc
     3541 tttgcttttc ttcttggcac tacttttatg tcactattat cttgtattac tactgcccct
     3601 tcacctttcc agaggagctt tgctggtcct ttccaaagtg gatttctgct gtccctgtaa
     3661 taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct
     3721 gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt
     3781 ttaaaattgt ggatgaatac tgccatttgt actgctgtct taagatgttc agcctgatct
     3841 cttacctgtc ctataatttt ctttaattct ttattcatag attctactac tccttgactt
     3901 tggggattgt agggaatttc caggtctgaa gatcctgctg cttgctgtgc ttttttctta
     3961 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     4021 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     4081 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     4141 aacaggccag tattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     4201 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     4261 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     4321 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     4381 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     4441 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     4501 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     4561 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     4621 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     4681 tccccagtcc cgcccaggcc acgcctccca tggtaatagc gatgactaat acgtagatgt
     4741 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     4801 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     4861 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     4921 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     4981 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     5041 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga cccaggtggc
     5101 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5161 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5221 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5281 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5341 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5401 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5461 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5521 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5581 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5641 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5701 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5761 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5821 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5881 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5941 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6001 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6061 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6121 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6181 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6241 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6301 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6361 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6421 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6481 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6541 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6601 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6661 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6721 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6781 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6841 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6901 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6961 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7021 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7081 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     7141 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     7201 ctgcaataaa caagttagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     7261 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     7321 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     7381 taggggcggg actatggttg ctgactaatt gagatgcgga tccgctggca cgacaggttt
     7441 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7501 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7561 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgg gctgcaggtc
     7621 gagggatctc cataagagaa gagggacagc tatgactggg agtagtcagg agaggaggaa
     7681 aaatctggct agtaaaacat gtaaggaaaa ttttagggat gttaaagaaa aaaataacac
     7741 aaaacaaaat at
//
