LOCUS       sequence-488179-        7314 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_159, derived from human fusion protein RABGAP1L_BCAS3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7314)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7314)
  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_159
FEATURES             Location/Qualifiers
     source          1..7314
                     /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(847..1557,1558..1560,1561..1563))
                     /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(889..910)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1238..1257
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1497..1518
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1560..1569
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1645..1848)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1674..1694)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1849..2152
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2409..2453)
                     /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    2638..2871
                     /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    3231..3348
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3840..3965
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4012..4269
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4271..4650
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4681..4785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4786..4854,4855..5646)
                     /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(5004..5023)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5817..6405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6306..6325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6559..6576
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6644..6778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6668..6687)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6722..6741
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6784..6979)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6798..6933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7038..7059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7074..7079,7080..7097,7098..7104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7112..7128
                     /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     7117..7139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7136..7152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7136..7152
                     /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 caaagaggga agtggcgaca aacgcgggtt
      601 ccctgggtta ttgttggtaa aattgttata gctgtcttgc gagtttaact tgccatgcag
      661 tggagagccc gagggagaac tgctaagtcc aggtactgga ctgcagcgtc caccttgctt
      721 tgaagtcagc tcttgttcga tttcctctaa cccagcagac ttttgaaagc ggctcatgcg
      781 attaaccacg cgcggactca tgtgccggga agcggccgca cttcccccgg agcctccgct
      841 tccgcccttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
      901 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact gggtgctcag
      961 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     1021 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     1081 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     1141 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     1201 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     1261 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     1321 ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac
     1381 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     1441 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     1501 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     1561 catggtggcg accggtggat cccgggcccg cggtaccgtc gactgcagaa ttcggttcac
     1621 taaacgagcc cggttcacta aacgagctct gcttatatag acctcccacc gtacacgcct
     1681 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     1741 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     1801 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     1861 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     1921 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     1981 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     2041 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     2101 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     2161 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac tagcacgcgt
     2221 tcgaagcttg agctcggtcc cctcggggtt gggaggtggg tctgaaacga taatggtgaa
     2281 tatccctgcc taactctatt cactatagaa agtacagcaa aaactattct taaacctacc
     2341 aagcctccta ctatcattat gaataatttt atataccaca gccaatttgt tatgttaaac
     2401 caattccaca aacttgccca tttatctaat tccaataatt cttgttcatt cttttcttgc
     2461 tggttttgcg attcttcaat taaggagtgt attaagcttg tgtaattgtt aatttctctg
     2521 tcccactcca tccaggtcgt gtgattccaa atctgttcca gagatttatt actccaacta
     2581 gcattccaag gcacagcagt ggtgcaaatg agttttccag agcaacccca aatccccagg
     2641 agctgttgat cctttaggta tctttccaca gccaggattc ttgcctggag ctgcttgatg
     2701 ccccagactg tgagttgcaa cagatgctgt tgcgcctcaa tagccctcag caaattgttc
     2761 tgctgctgca ctataccaga caataattgt ctggcctgta ccgtcagcgt cattgacgct
     2821 gcgcccatag tgcttcctgc tgctcccaag aacccaagga acaaagctcc tattcccact
     2881 gctctttttt ctctctgcac cactcttctc tttgccttgg tgggtgctac tcctaatggt
     2941 tcaattttta ctactttata tttatataat tcacttctcc aattgtccct catatctcct
     3001 ccttatggtg ttttactaaa cttttccatg ttctaatcct catcctgtct acttgccaca
     3061 caatcatcac ctgccatctg ttttccataa tccctaatga tctttgcttt tcttcttggc
     3121 actactttta tgtcactatt atcttgtatt actactgccc cttcaccttt ccagaggagc
     3181 tttgctggtc ctttccaaag tggatttctg ctgtccctgt aataaacccg aaaattttga
     3241 atttttgtaa tttgtttttg taattcttta gtttgtatgt ctgttgctat tatgtctact
     3301 attctttccc ctgcactgta ccccccaatc cccccttttc ttttaaaatt gtggatgaat
     3361 actgccattt gtactgctgt cttaagatgt tcagcctgat ctcttacctg tcctataatt
     3421 ttctttaatt ctttattcat agattctact actccttgac tttggggatt gtagggaatt
     3481 tccaggtctg aagatcctgc tgcttgctgt gcttttttct tacttttgtt ttgctcttcc
     3541 tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg cacacaatag
     3601 agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg aagggatggt
     3661 tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc agtattaact
     3721 gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata ttttttcttt
     3781 ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg cttaatactg
     3841 acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt tggcgtactc
     3901 accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga
     3961 gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca ctgctagaga
     4021 ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac
     4081 gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc
     4141 ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacagg
     4201 caaaaagcag ctgcttatat gcaggatctg agggctcgcc actccccagt cccgcccagg
     4261 ccacgcctcc catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     4321 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     4381 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     4441 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     4501 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     4561 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     4621 actattaata actagtcaat aatcaatgtc gacccaggtg gcacttttcg gggaaatgtg
     4681 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4741 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4801 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4861 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4921 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4981 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5041 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5101 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5161 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5221 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5281 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5341 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5401 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5461 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5521 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5581 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5641 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5701 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5761 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5821 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5881 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5941 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6001 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6061 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6121 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6181 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6241 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6301 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6361 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6421 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6481 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6541 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6601 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gcggatccag acatgataag
     6661 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     6721 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttag
     6781 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     6841 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     6901 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     6961 tgctgactaa ttgagatgcg gatccgctgg cacgacaggt ttcccgactg gaaagcgggc
     7021 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     7081 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     7141 aacagctatg accatgatta cgccaagctt gggctgcagg tcgagggatc tccataagag
     7201 aagagggaca gctatgactg ggagtagtca ggagaggagg aaaaatctgg ctagtaaaac
     7261 atgtaaggaa aattttaggg atgttaaaga aaaaaataac acaaaacaaa atat
//
