LOCUS       sequence-490787-        7791 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_196, derived from human fusion protein TMPRSS2_ETV1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7791)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7791)
  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_196
FEATURES             Location/Qualifiers
     source          1..7791
                     /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(1324..2034,2035..2037,2038..2040))
                     /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(1366..1387)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1715..1734
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1974..1995
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2037..2046
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2122..2325)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2151..2171)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2326..2629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2886..2930)
                     /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    3115..3348
                     /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    3708..3825
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4317..4442
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4489..4746
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4748..5127
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5158..5262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5263..5331,5332..6123)
                     /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(5481..5500)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6294..6882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6783..6802
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7036..7053
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7121..7255
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7145..7164)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7199..7218
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7261..7456)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7275..7410
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7515..7536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7551..7556,7557..7574,7575..7581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7589..7605
                     /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     7594..7616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7613..7629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7613..7629
                     /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 acccttttca aacatgcatc cctcggtacg
      601 gctcgggtgg gctaaaaagc cctcttgacg catataaata ctatggcacg acgggacttc
      661 actatcatag gcaaaatcgc gaggctcctg cttaatcatt aacggtggag gaaaggactg
      721 acttgccgct gatccgacca ttgtattatg ttcatacaca ggatcatggt actcttgttt
      781 aaatccctgt ggggggaacg ggatgttagg ttcactcatt tggcgttggt acatcggacg
      841 tccctcgcgc ggcatagtgg gcagcggtgg gaatgaatta catggctctg acaactgacg
      901 acgaaagcga tgatccatcg gatatgacga gtcaggaata ctttgggatg gtggcagatg
      961 ggctggaaaa gcgcgatcgg gctttggtgt gtgggtgctg ttaggactgg cgtggtgcag
     1021 tggtgatact ggcgtgctgg agggggtagg tgggttagac gggcgcatgc ctacttgcgg
     1081 tttttgatca taagcgctga cgttatacag gcatttctcc ccataactga atttgaacgg
     1141 ttgttcctgg ctgcatgccg aagagatctc agaacaggga gaatggggct ccttcttaat
     1201 tttcaacggt aatccgtgga aagctacagc agccccacgc tcctcgcatc ctgattctcc
     1261 acctgggggg gcagggggca tccgggaagc ggccgcactt cccccggagc ctccgcttcc
     1321 gcccttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     1381 gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggg tgctcaggta
     1441 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     1501 ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc
     1561 gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt
     1621 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1681 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1741 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1801 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     1861 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
     1921 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1981 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     2041 ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac tgcagaattc ggttcactaa
     2101 acgagcccgg ttcactaaac gagctctgct tatatagacc tcccaccgta cacgcctacc
     2161 gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg
     2221 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
     2281 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat
     2341 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     2401 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     2461 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     2521 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     2581 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     2641 atatgggcta tgaactaatg accccgtaat tgattactat taataactag cacgcgttcg
     2701 aagcttgagc tcggtcccct cggggttggg aggtgggtct gaaacgataa tggtgaatat
     2761 ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa acctaccaag
     2821 cctcctacta tcattatgaa taattttata taccacagcc aatttgttat gttaaaccaa
     2881 ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt ttcttgctgg
     2941 ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat ttctctgtcc
     3001 cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact ccaactagca
     3061 ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat ccccaggagc
     3121 tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg cttgatgccc
     3181 cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa attgttctgc
     3241 tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat tgacgctgcg
     3301 cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat tcccactgct
     3361 cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc taatggttca
     3421 atttttacta ctttatattt atataattca cttctccaat tgtccctcat atctcctcct
     3481 tatggtgttt tactaaactt ttccatgttc taatcctcat cctgtctact tgccacacaa
     3541 tcatcacctg ccatctgttt tccataatcc ctaatgatct ttgcttttct tcttggcact
     3601 acttttatgt cactattatc ttgtattact actgcccctt cacctttcca gaggagcttt
     3661 gctggtcctt tccaaagtgg atttctgctg tccctgtaat aaacccgaaa attttgaatt
     3721 tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat gtctactatt
     3781 ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg gatgaatact
     3841 gccatttgta ctgctgtctt aagatgttca gcctgatctc ttacctgtcc tataattttc
     3901 tttaattctt tattcataga ttctactact ccttgacttt ggggattgta gggaatttcc
     3961 aggtctgaag atcctgctgc ttgctgtgct tttttcttac ttttgttttg ctcttcctct
     4021 atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac acaatagagg
     4081 gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag ggatggttgt
     4141 agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagt attaactgcg
     4201 aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt tttctttccc
     4261 cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt aatactgacg
     4321 ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg cgtactcacc
     4381 agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag
     4441 agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg ctagagattt
     4501 tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg
     4561 cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta
     4621 gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca gtacaggcaa
     4681 aaagcagctg cttatatgca ggatctgagg gctcgccact ccccagtccc gcccaggcca
     4741 cgcctcccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     4801 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     4861 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     4921 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     4981 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     5041 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     5101 attaataact agtcaataat caatgtcgac ccaggtggca cttttcgggg aaatgtgcgc
     5161 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5221 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5281 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5341 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5401 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5461 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5521 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5581 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5641 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5701 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5761 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5821 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5881 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5941 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6001 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6061 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6121 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6181 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6241 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6301 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6361 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6421 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6481 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6541 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6601 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6661 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6721 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6781 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6841 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6901 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6961 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7021 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     7081 ccgcctctcc ccgcgcgttg gccgattcat taatgcagcg gatccagaca tgataagata
     7141 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     7201 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttagctt
     7261 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     7321 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     7381 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     7441 tgactaattg agatgcggat ccgctggcac gacaggtttc ccgactggaa agcgggcagt
     7501 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     7561 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7621 agctatgacc atgattacgc caagcttggg ctgcaggtcg agggatctcc ataagagaag
     7681 agggacagct atgactggga gtagtcagga gaggaggaaa aatctggcta gtaaaacatg
     7741 taaggaaaat tttagggatg ttaaagaaaa aaataacaca aaacaaaata t
//
