LOCUS       sequence-488215-        7500 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_197, derived from human fusion protein TMPRSS2_ETV1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7500)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7500)
  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_197
FEATURES             Location/Qualifiers
     source          1..7500
                     /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(1033..1743,1744..1746,1747..1749))
                     /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(1075..1096)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1424..1443
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1683..1704
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1746..1755
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1831..2034)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1860..1880)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2035..2338
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2595..2639)
                     /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    2824..3057
                     /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    3417..3534
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4026..4151
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4198..4455
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4457..4836
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4867..4971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4972..5040,5041..5832)
                     /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(5190..5209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6003..6591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6492..6511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6745..6762
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6830..6964
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6854..6873)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6908..6927
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6970..7165)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6984..7119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7224..7245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7260..7265,7266..7283,7284..7290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7298..7314
                     /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     7303..7325
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7322..7338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7322..7338
                     /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 atcgcgaggc tcctgcttaa tcattaacgg
      601 tggaggaaag gactgacttg ccgctgatcc gaccattgta ttatgttcat acacaggatc
      661 atggtactct tgtttaaatc cctgtggggg gaacgggatg ttaggttcac tcatttggcg
      721 ttggtacatc ggacgtccct cgcgcggcat agtgggcagc ggtgggaatg aattacatgg
      781 ctctgacaac tgacgacgaa agcgatgatc catcggatat gacgagtcag gaatactttg
      841 ggatggtggc agatgggctg gaaaagcgcg atcgggcttt ggtgtgtggg tgctgttagg
      901 actggcgtgg tgcagtggtg atactggcgt gctggagggg gtaggtgggt tagacgggcg
      961 catgcctact tgcggttttt gatcataagc ccgggaagcg gccgcacttc ccccggagcc
     1021 tccgcttccg cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     1081 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactgggt
     1141 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     1201 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac
     1261 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta
     1321 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     1381 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     1441 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     1501 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt
     1561 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     1621 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     1681 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     1741 gctcaccatg gtggcgaccg gtggatcccg ggcccgcggt accgtcgact gcagaattcg
     1801 gttcactaaa cgagcccggt tcactaaacg agctctgctt atatagacct cccaccgtac
     1861 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     1921 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     1981 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
     2041 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     2101 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     2161 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     2221 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     2281 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     2341 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactagc
     2401 acgcgttcga agcttgagct cggtcccctc ggggttggga ggtgggtctg aaacgataat
     2461 ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa
     2521 cctaccaagc ctcctactat cattatgaat aattttatat accacagcca atttgttatg
     2581 ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt
     2641 tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt
     2701 tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc
     2761 caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc
     2821 cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc ctggagctgc
     2881 ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa
     2941 ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt cagcgtcatt
     3001 gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa agctcctatt
     3061 cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg tgctactcct
     3121 aatggttcaa tttttactac tttatattta tataattcac ttctccaatt gtccctcata
     3181 tctcctcctt atggtgtttt actaaacttt tccatgttct aatcctcatc ctgtctactt
     3241 gccacacaat catcacctgc catctgtttt ccataatccc taatgatctt tgcttttctt
     3301 cttggcacta cttttatgtc actattatct tgtattacta ctgccccttc acctttccag
     3361 aggagctttg ctggtccttt ccaaagtgga tttctgctgt ccctgtaata aacccgaaaa
     3421 ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt tgctattatg
     3481 tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt aaaattgtgg
     3541 atgaatactg ccatttgtac tgctgtctta agatgttcag cctgatctct tacctgtcct
     3601 ataattttct ttaattcttt attcatagat tctactactc cttgactttg gggattgtag
     3661 ggaatttcca ggtctgaaga tcctgctgct tgctgtgctt ttttcttact tttgttttgc
     3721 tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct ttgatgcaca
     3781 caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc tgtctgaagg
     3841 gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa caggccagta
     3901 ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa tttatatttt
     3961 ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct cccccgctta
     4021 atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa aatttttggc
     4081 gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg
     4141 cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc
     4201 tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca
     4261 acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg
     4321 ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag
     4381 tacaggcaaa aagcagctgc ttatatgcag gatctgaggg ctcgccactc cccagtcccg
     4441 cccaggccac gcctcccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     4501 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     4561 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     4621 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     4681 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     4741 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     4801 ttgattacta ttaataacta gtcaataatc aatgtcgacc caggtggcac ttttcgggga
     4861 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4921 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4981 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5041 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5101 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5161 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5221 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5281 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5341 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5401 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5461 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5521 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5581 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5641 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5701 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5761 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5821 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5881 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5941 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6001 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6061 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6121 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6661 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6721 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6781 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcgg atccagacat
     6841 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     6901 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     6961 agttagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     7021 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     7081 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     7141 tatggttgct gactaattga gatgcggatc cgctggcacg acaggtttcc cgactggaaa
     7201 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7261 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7321 acaggaaaca gctatgacca tgattacgcc aagcttgggc tgcaggtcga gggatctcca
     7381 taagagaaga gggacagcta tgactgggag tagtcaggag aggaggaaaa atctggctag
     7441 taaaacatgt aaggaaaatt ttagggatgt taaagaaaaa aataacacaa aacaaaatat
//
