LOCUS       sequence-488181-        7383 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_161, derived from human fusion protein RABGAP1L_BCAS3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7383)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7383)
  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_161
FEATURES             Location/Qualifiers
     source          1..7383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(363..382)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    428..483
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(482..501)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(844..860)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(844..860)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(857..879)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    868..884
                     /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)."
     promoter        complement(join(892..898,899..916,917..922))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    937..958
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1017..1212
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1063..1198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1218..1352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1255..1274)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1309..1328
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1420..1437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1591..2179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1671..1690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2350..3141,3142..3210))
                     /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     2973..2992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3211..3315)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3346..3725
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             3727..3984
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4031..4156
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4648..4765
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5125..5358
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5543..5587
                     /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"
     enhancer        5844..6147
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6148..6351
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6302..6322
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6427..6436
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6433..6435,6436..6438,6439..7149)
                     /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(6478..6499)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6739..6758)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7086..7107
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aaaaaaaaac aatgtcagca accatctgtc cgcgagcaac ctaactccaa caaagcgtgt
       61 gtacgtgatg gtgggcgtac ctcagcacgc ggcaaacatc gcgacagcga atgaaccggt
      121 tctagataag attacggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
      181 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaaa gaagacaaga
      241 tctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
      301 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttcagctgct cgagctagca
      361 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
      421 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
      481 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
      541 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
      601 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
      661 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
      721 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
      781 atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc
      841 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
      901 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      961 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
     1021 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     1081 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     1141 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     1201 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1261 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1321 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
     1381 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1441 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1501 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1561 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1621 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1681 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1741 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1801 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1861 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1921 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1981 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2041 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2101 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2161 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2221 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2281 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2341 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2401 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2461 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2521 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2581 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2641 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2701 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2761 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2821 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2881 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2941 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3001 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3061 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3121 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3181 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3241 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3301 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgattattga
     3361 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     3421 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     3481 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     3541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     3601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     3661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     3721 ccatgggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata
     3781 agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
     3841 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     3901 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     3961 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
     4021 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     4081 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     4141 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     4201 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     4261 gctagaacga ttcgcagtta atactggcct gttagaaaca tcagaaggct gtagacaaat
     4321 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     4381 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     4441 tttagacaag atagaggaag agcaaaacaa aagtaagaaa aaagcacagc aagcagcagg
     4501 atcttcagac ctggaaattc cctacaatcc ccaaagtcaa ggagtagtag aatctatgaa
     4561 taaagaatta aagaaaatta taggacaggt aagagatcag gctgaacatc ttaagacagc
     4621 agtacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
     4681 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     4741 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagaa atccactttg
     4801 gaaaggacca gcaaagctcc tctggaaagg tgaaggggca gtagtaatac aagataatag
     4861 tgacataaaa gtagtgccaa gaagaaaagc aaagatcatt agggattatg gaaaacagat
     4921 ggcaggtgat gattgtgtgg caagtagaca ggatgaggat tagaacatgg aaaagtttag
     4981 taaaacacca taaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     5041 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     5101 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     5161 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     5221 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     5281 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     5341 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     5401 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     5461 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     5521 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     5581 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     5641 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     5701 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     5761 gagctcaagc ttcgaacgcg tgctagttat taatagtaat caattacggg gtcattagtt
     5821 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     5881 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     5941 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6001 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     6061 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     6121 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     6181 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     6241 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     6301 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
     6361 aaccgggctc gtttagtgaa ccgaattctg cagtcgacgg taccgcgggc ccgggatcca
     6421 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     6481 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     6541 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     6601 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     6661 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     6721 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     6781 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     6841 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     6901 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     6961 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     7021 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     7081 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     7141 ctgtacaagg gcggaagcgg aggctccggg ggaagtgcgg ccgcttcccg gtttggtctg
     7201 cgtcacatgt cgagcatgga acacaccgag gagggattgc gtgaacgttt agccgatgcg
     7261 atggcagagt ccccttcgcg tgatgtagta gggagtggta cagatacggc gctggacgtg
     7321 gctgttaaga ctttcccgcc agagcgccat gttgctgtaa aatgttttgg caaaaagaag
     7381 ggg
//
