LOCUS       sequence-488216-        7926 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_198, derived from human fusion protein TMPRSS2_ETV4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7926)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7926)
  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_198
FEATURES             Location/Qualifiers
     source          1..7926
                     /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(1459..2169,2170..2172,2173..2175))
                     /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(1501..1522)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1850..1869
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2109..2130
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2172..2181
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2257..2460)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2286..2306)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2461..2764
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3021..3065)
                     /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    3250..3483
                     /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    3843..3960
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4452..4577
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4624..4881
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4883..5262
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5293..5397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5398..5466,5467..6258)
                     /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(5616..5635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6429..7017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6918..6937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7171..7188
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7256..7390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7280..7299)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7334..7353
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7396..7591)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7410..7545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7650..7671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7686..7691,7692..7709,7710..7716)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7724..7740
                     /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     7729..7751
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7748..7764
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7748..7764
                     /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 cttaatgact actcctgcac cggggtagcg
      601 atgtccattc acaccgcctt gatccacggc cggttgtcct gcctgttcat aaagagggtc
      661 atggtactct tgtttgaaag attgctgcgg atacggcggg catggctcag ataattggtg
      721 ctgatagggt gcaggcaggg gttcgcgccc tccaccttga gatgtgaacg agtggcaaat
      781 gtccagaggc tgttgaaaga ctgaggagtg ttcgcccaga tacccgtgtc ccggatgagg
      841 ttgactagtt cccgaggaac gaaggaaatt gcgctgttca gcacgaggaa atggttgaag
      901 aggactttgg cccaaagcac ctggtgctgg acttttgatt gcaatttggc gaggggggtc
      961 atacgcggag gagtaaaggc actgctcacc atgatggtag ggcagtggag gtttgcggct
     1021 acaggaaaga gctgggtctg tacgaggaga ttgtggttct ttcttgatgc gcgtggttgg
     1081 gctgtgaaat gctaagtttt cactatgaaa gtccggtacg aactgctcgt ccgaatcggg
     1141 gacctgggct tcagcaagcc aggtttcctg gaagtggctc aggtcttgga acaagtcctc
     1201 ggagtccaat gggggcaatg atcctgggtc catcagtttg cccaagggtc caatcagggc
     1261 ctcgcgcagg gacccgtttc caggtgactt gctatttaag gccattttcg aattatctac
     1321 cgcatctaac aacgaaagat aacgcgaatg ttcgatatga cccgcagccc cacgttcttc
     1381 acatccggac tctccgcccg gtggagccgg aggcatccgg gaagcggccg cacttccccc
     1441 ggagcctccg cttccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     1501 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga
     1561 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     1621 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     1681 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     1741 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     1801 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     1861 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     1921 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     1981 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     2041 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     2101 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     2161 gcccttgctc accatggtgg cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag
     2221 aattcggttc actaaacgag cccggttcac taaacgagct ctgcttatat agacctccca
     2281 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     2341 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     2401 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     2461 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     2521 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     2581 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     2641 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     2701 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     2761 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     2821 actagcacgc gttcgaagct tgagctcggt cccctcgggg ttgggaggtg ggtctgaaac
     2881 gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt
     2941 cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt
     3001 gttatgttaa accaattcca caaacttgcc catttatcta attccaataa ttcttgttca
     3061 ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg
     3121 ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta
     3181 ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc
     3241 caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg
     3301 agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc
     3361 agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc
     3421 gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct
     3481 cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct
     3541 actcctaatg gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc
     3601 ctcatatctc ctccttatgg tgttttacta aacttttcca tgttctaatc ctcatcctgt
     3661 ctacttgcca cacaatcatc acctgccatc tgttttccat aatccctaat gatctttgct
     3721 tttcttcttg gcactacttt tatgtcacta ttatcttgta ttactactgc cccttcacct
     3781 ttccagagga gctttgctgg tcctttccaa agtggatttc tgctgtccct gtaataaacc
     3841 cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct
     3901 attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa
     3961 ttgtggatga atactgccat ttgtactgct gtcttaagat gttcagcctg atctcttacc
     4021 tgtcctataa ttttctttaa ttctttattc atagattcta ctactccttg actttgggga
     4081 ttgtagggaa tttccaggtc tgaagatcct gctgcttgct gtgctttttt cttacttttg
     4141 ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga
     4201 tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc
     4261 tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg
     4321 ccagtattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta
     4381 tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc
     4441 cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt
     4501 tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga
     4561 gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc
     4621 cactgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     4681 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     4741 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     4801 acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg ccactcccca
     4861 gtcccgccca ggccacgcct cccatggtaa tagcgatgac taatacgtag atgtactgcc
     4921 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     4981 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     5041 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     5101 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     5161 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     5221 ccgtaattga ttactattaa taactagtca ataatcaatg tcgacccagg tggcactttt
     5281 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5341 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5401 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5461 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5521 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5581 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5641 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5701 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5761 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5821 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5881 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5941 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6001 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6061 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6121 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6181 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6241 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6301 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6361 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6421 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6481 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6541 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6601 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6661 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6721 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6781 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6841 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6901 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6961 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7021 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7081 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7141 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7201 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcggatcc
     7261 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     7321 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     7381 taaacaagtt agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     7441 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     7501 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     7561 cgggactatg gttgctgact aattgagatg cggatccgct ggcacgacag gtttcccgac
     7621 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7681 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7741 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgggctgca ggtcgaggga
     7801 tctccataag agaagaggga cagctatgac tgggagtagt caggagagga ggaaaaatct
     7861 ggctagtaaa acatgtaagg aaaattttag ggatgttaaa gaaaaaaata acacaaaaca
     7921 aaatat
//
