LOCUS       sequence-490788-        8064 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_199, derived from human fusion protein TMPRSS2_ETV4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8064)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8064)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_199
FEATURES             Location/Qualifiers
     source          1..8064
                     /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(1597..2307,2308..2310,2311..2313))
                     /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(1639..1660)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1988..2007
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2247..2268
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2310..2319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2395..2598)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2424..2444)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2599..2902
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3159..3203)
                     /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    3388..3621
                     /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    3981..4098
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4590..4715
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4762..5019
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5021..5400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5431..5535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5536..5604,5605..6396)
                     /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(5754..5773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6567..7155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7056..7075
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7309..7326
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7394..7528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7418..7437)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7472..7491
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7534..7729)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7548..7683
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7788..7809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7824..7829,7830..7847,7848..7854)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7862..7878
                     /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     7867..7889
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7886..7902
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7886..7902
                     /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 cgggcgtaac ggtttctcat agccgtatcc
      601 cattgcgcca tcacctgggg aaggtcccga gaatccttct gtatgcaggt acatggacgc
      661 acagcccgtt acatccgaat cgtacgcaaa atcggtctgc tcttgcttaa tgactactcc
      721 tgcaccgggg tagcgatgtc cattcacacc gccttgatcc acggccggtt gtcctgcctg
      781 ttcataaaga gggtcatggt actcttgttt gaaagattgc tgcggatacg gcgggcatgg
      841 ctcagataat tggtgctgat agggtgcagg caggggttcg cgccctccac cttgagatgt
      901 gaacgagtgg caaatgtcca gaggctgttg aaagactgag gagtgttcgc ccagataccc
      961 gtgtcccgga tgaggttgac tagttcccga ggaacgaagg aaattgcgct gttcagcacg
     1021 aggaaatggt tgaagaggac tttggcccaa agcacctggt gctggacttt tgattgcaat
     1081 ttggcgaggg gggtcatacg cggaggagta aaggcactgc tcaccatgat ggtagggcag
     1141 tggaggtttg cggctacagg aaagagctgg gtctgtacga ggagattgtg gttctttctt
     1201 gatgcgcgtg gttgggctgt gaaatgctaa gttttcacta tgaaagtccg gtacgaactg
     1261 ctcgtccgaa tcggggacct gggcttcagc aagccaggtt tcctggaagt ggctcaggtc
     1321 ttggaacaag tcctcggagt ccaatggggg caatgatcct gggtccatca gtttgcccaa
     1381 gggtccaatc agggcctcgc gcagggaccc gtttccaggt gacttgctat ttaaggccat
     1441 tttcgaatta tctaccgcat ctaacaacga aagataacgc gaatgttcga tatgacccgc
     1501 agccccacgt tcttcacatc cggactctcc gcccggtgga gccggaggca ttccccggga
     1561 agcggccgca cttcccccgg agcctccgct tccgcccttg tacagctcgt ccatgccgag
     1621 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
     1681 gtctttgctc agcttggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc
     1741 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat
     1801 gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata
     1861 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     1921 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
     1981 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
     2041 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa
     2101 gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     2161 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     2221 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     2281 cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtggat cccgggcccg
     2341 cggtaccgtc gactgcagaa ttcggttcac taaacgagcc cggttcacta aacgagctct
     2401 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     2461 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     2521 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     2581 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     2641 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     2701 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     2761 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     2821 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     2881 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     2941 aattgattac tattaataac tagcacgcgt tcgaagcttg agctcggtcc cctcggggtt
     3001 gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt cactatagaa
     3061 agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat gaataatttt
     3121 atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca tttatctaat
     3181 tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat taaggagtgt
     3241 attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt gtgattccaa
     3301 atctgttcca gagatttatt actccaacta gcattccaag gcacagcagt ggtgcaaatg
     3361 agttttccag agcaacccca aatccccagg agctgttgat cctttaggta tctttccaca
     3421 gccaggattc ttgcctggag ctgcttgatg ccccagactg tgagttgcaa cagatgctgt
     3481 tgcgcctcaa tagccctcag caaattgttc tgctgctgca ctataccaga caataattgt
     3541 ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc tgctcccaag
     3601 aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac cactcttctc
     3661 tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata tttatataat
     3721 tcacttctcc aattgtccct catatctcct ccttatggtg ttttactaaa cttttccatg
     3781 ttctaatcct catcctgtct acttgccaca caatcatcac ctgccatctg ttttccataa
     3841 tccctaatga tctttgcttt tcttcttggc actactttta tgtcactatt atcttgtatt
     3901 actactgccc cttcaccttt ccagaggagc tttgctggtc ctttccaaag tggatttctg
     3961 ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg taattcttta
     4021 gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta ccccccaatc
     4081 cccccttttc ttttaaaatt gtggatgaat actgccattt gtactgctgt cttaagatgt
     4141 tcagcctgat ctcttacctg tcctataatt ttctttaatt ctttattcat agattctact
     4201 actccttgac tttggggatt gtagggaatt tccaggtctg aagatcctgc tgcttgctgt
     4261 gcttttttct tacttttgtt ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct
     4321 tttatctcta tcctttgatg cacacaatag agggttgcta ctgtattata taatgatcta
     4381 agttcttctg atcctgtctg aagggatggt tgtagctgtc ccagtatttg tctacagcct
     4441 tctgatgttt ctaacaggcc agtattaact gcgaatcgtt ctagctccct gcttgcccat
     4501 actatatgtt ttaatttata ttttttcttt ccccctggcc ttaaccgaat tttttcccat
     4561 cgcgatctaa ttctcccccg cttaatactg acgctctcgc acccatctct ctccttctag
     4621 cctccgctag tcaaaatttt tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg
     4681 cgcgcttcag caagccgagt cctgcgtcga gagagctcct ctggtttccc tttcgctttc
     4741 aagtccctgt tcgggcgcca ctgctagaga ttttccacac tgactaaaag ggtctgaggg
     4801 atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca
     4861 agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga
     4921 tctggtctaa ccagagagac ccagtacagg caaaaagcag ctgcttatat gcaggatctg
     4981 agggctcgcc actccccagt cccgcccagg ccacgcctcc catggtaata gcgatgacta
     5041 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     5101 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     5161 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     5221 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     5281 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     5341 ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc
     5401 gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5461 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5521 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5581 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5641 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5701 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5761 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5821 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5881 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5941 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6001 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6061 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6121 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6181 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6241 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6301 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6361 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6421 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6481 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6541 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6601 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6661 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6721 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6781 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6841 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6901 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6961 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7021 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7081 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7141 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7201 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7261 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7321 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     7381 cattaatgca gcggatccag acatgataag atacattgat gagtttggac aaaccacaac
     7441 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     7501 aaccattata agctgcaata aacaagttag ctttttgcaa aagcctaggc ctccaaaaaa
     7561 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     7621 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     7681 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgcg gatccgctgg
     7741 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     7801 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7861 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
     7921 gggctgcagg tcgagggatc tccataagag aagagggaca gctatgactg ggagtagtca
     7981 ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa aattttaggg atgttaaaga
     8041 aaaaaataac acaaaacaaa atat
//
