LOCUS       sequence-488214-        7578 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_195, derived from human fusion protein TMPRSS2_ERG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7578)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7578)
  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_195
FEATURES             Location/Qualifiers
     source          1..7578
                     /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(1111..1821,1822..1824,1825..1827))
                     /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(1153..1174)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1502..1521
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1761..1782
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1824..1833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1909..2112)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1938..1958)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2113..2416
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2673..2717)
                     /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    2902..3135
                     /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    3495..3612
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4104..4229
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4276..4533
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4535..4914
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4945..5049
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5050..5118,5119..5910)
                     /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(5268..5287)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6081..6669
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6570..6589
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6823..6840
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6908..7042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6932..6951)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6986..7005
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7048..7243)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7062..7197
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7302..7323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7338..7343,7344..7361,7362..7368)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7376..7392
                     /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     7381..7403
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7400..7416
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7400..7416
                     /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 gtcagcggga acaatcacgc gacgctcatt
      601 agttgtcatg ttcggtggag gcatgtgttt ctcctccatg taggacccgt agttcatccc
      661 taccgtatcc ggactgccaa ccattttgcc tcctttggct acgctacact catcgggcga
      721 attgcgtgac ccattaacct gcgatggatt gcattccatt ttgatagtga cacgtgcggg
      781 gggctgagac aaccagtctt gctgcgggac acgcggcgac attttacttg tctgcccata
      841 atccgacgac gaagatgctg tcatttcggt tttcgctaaa tgcggtgtcc cataagcaca
      901 ctcaaataag gattgatcct cggagactac actcagagct tcgctattca gtgccatctt
      961 actattgtca acggcatcca gcagagacaa ataacggctg tgttcaatat gaccagctgc
     1021 accgcgttcc tcacatcctg actcgccgcc aggcggagcg ggaggcatcc gggaagcggc
     1081 cgcacttccc ccggagcctc cgcttccgcc cttgtacagc tcgtccatgc cgagagtgat
     1141 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
     1201 gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
     1261 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg
     1321 gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt
     1381 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     1441 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     1501 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
     1561 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg
     1621 cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     1681 gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
     1741 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     1801 gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt ggatcccggg cccgcggtac
     1861 cgtcgactgc agaattcggt tcactaaacg agcccggttc actaaacgag ctctgcttat
     1921 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     1981 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     2041 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     2101 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     2161 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     2221 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     2281 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     2341 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     2401 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     2461 ttactattaa taactagcac gcgttcgaag cttgagctcg gtcccctcgg ggttgggagg
     2521 tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca
     2581 gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac
     2641 cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat
     2701 aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag
     2761 cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt
     2821 tccagagatt tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt
     2881 ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg
     2941 attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc
     3001 tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc
     3061 tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca
     3121 aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc
     3181 ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata taattcactt
     3241 ctccaattgt ccctcatatc tcctccttat ggtgttttac taaacttttc catgttctaa
     3301 tcctcatcct gtctacttgc cacacaatca tcacctgcca tctgttttcc ataatcccta
     3361 atgatctttg cttttcttct tggcactact tttatgtcac tattatcttg tattactact
     3421 gccccttcac ctttccagag gagctttgct ggtcctttcc aaagtggatt tctgctgtcc
     3481 ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc tttagtttgt
     3541 atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc aatcccccct
     3601 tttcttttaa aattgtggat gaatactgcc atttgtactg ctgtcttaag atgttcagcc
     3661 tgatctctta cctgtcctat aattttcttt aattctttat tcatagattc tactactcct
     3721 tgactttggg gattgtaggg aatttccagg tctgaagatc ctgctgcttg ctgtgctttt
     3781 ttcttacttt tgttttgctc ttcctctatc ttgtctaaag cttccttggt gtcttttatc
     3841 tctatccttt gatgcacaca atagagggtt gctactgtat tatataatga tctaagttct
     3901 tctgatcctg tctgaaggga tggttgtagc tgtcccagta tttgtctaca gccttctgat
     3961 gtttctaaca ggccagtatt aactgcgaat cgttctagct ccctgcttgc ccatactata
     4021 tgttttaatt tatatttttt ctttccccct ggccttaacc gaattttttc ccatcgcgat
     4081 ctaattctcc cccgcttaat actgacgctc tcgcacccat ctctctcctt ctagcctccg
     4141 ctagtcaaaa tttttggcgt actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct
     4201 tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt tccctttcgc tttcaagtcc
     4261 ctgttcgggc gccactgcta gagattttcc acactgacta aaagggtctg agggatctct
     4321 agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt
     4381 attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt
     4441 ctaaccagag agacccagta caggcaaaaa gcagctgctt atatgcagga tctgagggct
     4501 cgccactccc cagtcccgcc caggccacgc ctcccatggt aatagcgatg actaatacgt
     4561 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     4621 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     4681 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     4741 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     4801 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     4861 gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgtcgaccca
     4921 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4981 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5041 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5101 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5161 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5221 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5281 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5341 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5401 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5461 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5521 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5581 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5641 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5701 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5761 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5821 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5881 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5941 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6001 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6061 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6121 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6181 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6241 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6301 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6361 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6421 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6481 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6541 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6601 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6661 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6721 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6781 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6841 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6901 tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     6961 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     7021 tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     7081 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     7141 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     7201 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcggatccg ctggcacgac
     7261 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7321 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7381 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgggctg
     7441 caggtcgagg gatctccata agagaagagg gacagctatg actgggagta gtcaggagag
     7501 gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa
     7561 taacacaaaa caaaatat
//
